Profiling of derived-hepatocyte progenitors from induced pluripotent stem cells using nanoCAGE promoter analysis
2019
Lereau Bernier, Myriam | Poulain, Stéphane | Tauran, Yannick | Danoy, Mathieu | Shinohara, Marie | Kimura, Keiichi | Segard, Bertrand David | Kato, Sachi | Kido, Taketomo | Miyajima, Atsushi | Sakai, Yasuyuki | Plessy, Charles | Leclerc, Eric
Human induced pluripotent stem (hiPS) cells represent a potential source of human cells for regenerative and personalized medicine applications. It is envisioned that hiPS cells could be an efficient tool for disease modelling and drug therapy testings. However, the successful derivation of hiPS cells to mature hepatocytes remains a challenge as far as the current proposed protocols lead to immature patterns and are very expensive (due to growth factors cost). In this work, we have compared two protocols of differentiation to produce hepatocyte-like cells. The first protocol is the conventional protocol adapted from Si-Tayeb et al. in 2010. By changing the frequency of culture medium exchange, we reduced the use of growth factors in the second protocol by 40%. Both protocols lead to hepatocyte-like characteristics including albumin (ALB) and cytochrome P450/3A4 (CYP3A4) expression in cells. Albumin production reached 2500 ng/mL/day for both protocols. The transcription factor motif activity analysis via transcriptomic data obtained with the Cap Analysis Gene Expression (CAGE) technology confirmed a typical hepatic profile for both protocols (such as activation of HNF1 A and HNF1B promoters). However, specific patterns were also detected with higher mRNA levels of WT1, MTF1, HCFC1, and PGCα1 in protocol 2. The overall transcriptomic analysis also revealed a modulation of targets involved in glucose homeostasis, in cell remodeling, lipid pathways and chromatin processing. However, the alpha-fetoprotein (AFP) production remained very high and the function of CYP3A4 was weak in both protocols illustrating still an immature profile in both protocols.
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